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Food and Chemical Toxicology 44 (2006) 847–854 www.elsevier.com/locate/foodchemtox Degradation of polychlorinated
Food and Chemical Toxicology 44 (2006) 847–854 www.elsevier.com/locate/foodchemtox Degradation of polychlorinated

Food and Chemical Toxicology 44 (2006) 847–854

Food and Chemical Toxicology 44 (2006) 847–854 www.elsevier.com/locate/foodchemtox Degradation of polychlorinated

www.elsevier.com/locate/foodchemtox

Degradation of polychlorinated biphenyls (PCBs) by Staphylococcus xylosus in liquid media and meat mixture

F.L. Lea˜ es a,b , A.P. Daniel a , G.B. Mello c , V. Battisti c , S. Bogusz Jr. d , T. Emanuelli e , L.L.M. Fries e , I. Costabeber c, *

a Programa de Po´ s-graduac¸a˜ o em Cieˆ ncia e Tecnologia de Alimentos, Centro de Cieˆ ncias Rurais, Universidade Federal de Santa Maria, Camobi, Santa Maria, RS, CEP: 97105-900, Brazil b Unidade Sa˜ o Luiz Gonzaga, Universidade Estadual do Rio Grande do Sul, Sa˜ o Luiz Gonzaga, RS, CEP: 97800-000, Brazil c Departamento de Morfologia, Centro de Cieˆ ncias da Sau´ de, Universidade Federal de Santa Maria, Campus de Camobi, Santa Maria, RS, CEP: 97105-900, Brazil

d Departamento de Cieˆ ncias da Sau´ de, Universidade Regional do Noroeste do Rio Grande do Sul, Ijuı´ , RS, CEP: 98700-000, Brazil

e Departamento de Tecnologia e Cieˆ ncia dos Alimentos, Centro de Cieˆ ncias Rurais, Universidade Federal de Santa Maria, Camobi, Santa Maria, RS, CEP: 97105-900, Brazil

Received 24 June 2005; accepted 15 November 2005

Abstract

We investigated the growth of the meat starter Staphylococcus xylosus (10 4 cells mL 1 ) in liquid media containing 0.01 ppm of each polychlorinated biphenyls (PCBs 10, 28, 52, 138, 153, and 180) and its ability to degrade PCBs during 168 h of incubation in liquid media (10 4 cells mL 1 , 0.01 ppm of each PCB congener) and cured meat mixture (0.1% of meat starter, 1 lg g 1 fat of each PCB congener). PCBs did not affect the growth of the starter microorganism in nutritive (brain heart infusion, BHI) or mineral salts medium (MSM) when compared to control (no PCB). S. xylosus degraded some of the PCB congeners tested. PCBs 138 and 153 were degraded both in BHI (78% and 68%, respectively; p < 0.05) and in MSM (71% and 66%, respectively; p < 0.05), with maximum degradation being observed within 24 h. Highly significant negative exponential relationships was observed between incubation time and concentrations of PCB 28 and 180 in BHI, as well as for PCBs 52 and 180 in MSM. In the cured meat mixture highly significant negative exponential relationship was observed between incubation time and the concentration of PCB 10. These results indicate that although S. xylosus reduced residues of various PCB congeners in liquid media, it was less effective in cured meat. 2005 Elsevier Ltd. All rights reserved.

Keywords: Degradation; Polychlorinated biphenyls; Staphylococcus xylosus, Meat mixture

1. Introduction

Polychlorinated biphenyls (PCBs) are a class of 209 compounds with the molecular formula C 12 H 10 n Cl n , where 1 6 n 6 10 ( Fig. 1 ). They were commercially

Abbreviations: ANOVA, analysis of variance; BHI, brain heart infus- ion; p, p 0 -DDT, 1,1,1-trichloro-2,2-bis-chloro phenyl ethane; MSM, min- eral salt medium; PCBs, polychlorinated biphenyls. * Corresponding author. Tel.: +55 55 32209375; fax: +55 55 32208494. E-mail address: ijoni@smail.ufsm.br (I. Costabeber).

0278-6915/$ - see front matter 2005 Elsevier Ltd. All rights reserved.

doi:10.1016/j.fct.2005.11.008

produced as complex mixtures for various uses, being employed mainly as dielectric fluids in capacitors and transformers ( WHO, 1993; Penteado and Moreira Vaz, 2001 ). Their thermal and chemical stability, resistance to chemical corrosion, and general inertness have contributed to their persistence in the environment (Haluska et al., 1995; Wiegel and Wu, 2000; Trı´ska et al., 2004 ). PCBs are deposited onto plants and water consumed by animals and fish. Due to their lipophilic properties and minimal degradation, they are biomagnified up the multi- step food chain (Zuccato et al., 1999; De Vos et al., 2003;

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es et al. / Food and Chemical Toxicology 44 (2006) 847–854 Fig. 1. Chemical structure of

Fig. 1. Chemical structure of PCBs ( Wiegel and Wu, 2000). In PCBs, some or all of the 10 hydrogens bound to positions 2–6 and 2 0 –6 0 are substituted with chlorines.

Weiglas-Kuperus et al., 2004 ). Hence, the major source of non-occupational exposure to these contaminants is via food consumption, particularly fish, meat, and dairy prod- ucts ( Focant et al., 2002; Ahmed, 2003 ). It is estimated that meat and meat products contribute to 14–19% of the cumulative toxic effects of PCBs when compared to the other foodstuffs (Patandin et al., 1999 ). Some of the effects described after exposure to PCBs include neurotoxicity, immunotoxicity, carcinogenicity, as well as endocrine disruption ( Ross, 2004 ). Several methods for PCBs destruction in environmental matrixes have been suggested ( Kubatova et al., 2001 ). However, because classical remediation techniques (cata- lytic process, incineration, etc.) are generally expensive, and not always efficient, nowadays the research has been focused on the development of bioremediation processes ( Sierra et al., 2003; Andreoni et al., 2004 ). Therefore, many microbes and enrichment cultures have been obtained that are able to metabolize and utilize PCBs as carbon and/or energy sources under aerobic or anaerobic conditions ( Abraham et al., 2002 ). However, degradation is a very specific process and the growth of some microorganisms can even be inhibited by a xenobiotic ( Bayarri, 1997; Abou-Arab, 2002 ). Studies on the degradation of PCB residues in meat and meat products are very important because of their increas- ing rate of consumption worldwide. Nowadays, the use of starter cultures is very common to improve characteristics of meat products. Fermented sausage is a high-fat-content meat product obtained using starter cultures of microor- ganisms like Micrococcus aurantiaccus , Pediococcus cerevi- sae ( Abou-Arab, 2002 ), and Staphylococcus xylosus ( Stahnke, 1995 ). The possibility that these microorganisms could degrade PCB residues is of great interest for the meat industry because it implies in a reduction of the toxicolog- ical risks associated to residues of highly stable com- pounds. However, few studies have evaluated this possibility ( Bayarri, 1997 ). The present study was carried out to investigate the growth of the meat starter S. xylosus in liquid media con- taining PCBs and its ability to degrade PCBs in liquid media and meat mixture.

2. Material and methods

2.1. Materials

Meat starter culture Bactoferm S-SX (S. xylosus) was supplied by Chr. Hansen Ind. e Com. Ltda. (Valinhos, SP, Brasil). The microorganism was isolated from the commercial meat starter culture. The strains were stored in skim milk vials at 18 C until use. An actively growing broth culture was prepared by incubating the strain in brain heart infusion broth at 37 C for 18 h. This culture was diluted to a final concentration of 10 5 cells mL 1 that was used to inoculate the sterile media in the experiments with PCBs. The appropriate dilution was previously determined by counting colony forming units in Baird–Parker agar (Difco, Detroit, MI) at 37 C for 48 h. Culture media for the in vitro assays were the following: brain heart infusion broth (BHI, Merck, Darmstadt, Germany); mineral salts medium (MSM) prepared according to Bayarri et al. (1998) and Baird–Parker Agar (Merck, Darmstadt, Germany). n -Hexane, acetone and petroleum ether were obtained from Mal- linckrodt (Kentucky, USA). Anhydrous sodium sulfate and 60/100-mesh pesticide reagent grade Florisil were obtained from Sigma (St. Louis, MO, USA). Florisil was previously activated at 150 C/12 h and partially deactivated by adding 2% Milli-Q water before use. Standard PCB solutions of 2,6-dichlorobiphenyl, 2,4,4 0 -trichlorobi- phenyl, 2,2 0 ,5,5 0 -tetrachlorobiphenyl, 2,2 0 ,3,4,4 0 ,5 0 -hexachlorobiphenyl, 2,2 0 ,4,4 0 ,5,5 0 -hexachlorobiphenyl, and 2,2 0 ,3,4,4 0 ,5,5 0 -heptachlorobiphenyl (IUPAC no. 10, 28, 52, 138, 153, and 180, respectively) were obtained from SUPELCO Inc. (Bellefonte, PA, USA). All other reagents used were of analytical reagent grade.

2.2. Methods

2.2.1. In vitro experiments with starter microorganism

The in vitro study was performed in two different liquid media:

nutritive medium (BHI) and a mineral salts medium (MSM). Tubes containing 8.99 mL of the sterilized liquid media (BHI or MSM) were inoculated with 1 mL S. xylosus (10 5 cells mL 1 concentration, to yield an initial cell number of 10 4 cells mL 1 in the inoculated media) and spiked with 0.01 mL of acetone or a standard solution of PCB congeners no. 10, 28, 52, 138, 153, and 180 prepared in acetone, in which each compound was at 10 ppm (l g mL 1 ). Unspiked samples (liquid media) were used as control. Samples were withdrawn at the beginning of the incubation (0 h) and after 3, 6, 12, 24, 48, 72, 96, 120, 144, and 168 h of incubation at 30 C for bacteria count to investigate any inhibitory effect of PCBs. Bacteria count was recorded after incubation in Baird–Parker agar at 37 C for 48 h. The residues of PCBs remaining in the medium were extracted with hexane following the method described by Angulo (1998), with some modifications. The medium (10 mL) was shaken with hexane (25 mL) and sulfuric acid (20 mL) in a separatory funnel. The aqueous layer was removed and the organic layer was extracted four times with 5 mL of sulfuric acid. The organic layer was removed and filtered through anhy- drous sodium sulfate, evaporated to dryness in a rotary evaporator, dis- solved in 2 mL of n-hexane, and used for PCB determination.

2.2.2. Meat mixture experiments

For the investigation of PCBs degradation by S. xylosus in a meat mixture, the meat mixture (1.2 kg pork meat, 0.4 kg bovine meat, 0.4 kg bacon, 6 g cure salt (sodium nitrite and nitrate), 5 g sodium ascorbate, 50 g sodium chloride, 20 g sucrose, 10 g commercial spices, and 0.1% starter culture) was contaminated with PCB congeners (1 lg g 1 fat for each

congener) and their concentrations were evaluated during incubation at 30 C for 7 days. Samples were analyzed at the beginning of the incubation (0 h) and after 3, 6, 12, 24, 48, 72, 96, 120, 144, and 168 h to determine PCB residues. Unspiked samples were used as control and the study was performed in triplicate. The fat of meat products was extracted as described by Waliszewski et al. (1997), with some modifications. In brief, homogenized samples (50 g) were ground with a sufficient amount of

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anhydrous sodium sulfate to yield a dry mixture, transferred to a 50 · 1 cm i.d. chromatographic column and extracted with 120 mL petroleum ether. The extract was transferred to a 50 mL round-bottom flask and the solvent was eliminated in a rotary evaporator. The extract (1 g) was applied to a chromatographic column containing 15 g of florisil and anhydrous sodium sulfate, and eluted with 100 mL n-hexane to extract PCBs. The eluate was filtered through anhydrous sodium sulfate, evaporated to dryness in a rotary evaporator, dissolved in 2 mL n-hexane, and used for PCB determination. All glassware used was previously cleaned by the method of Angulo et al. (1996).

2.2.3. Analysis of degradation products Chromatographic analyses were performed with a Hewlett Packard model 6890 gas chromatograph equipped with a 63 Ni electron capture

micro detector (GC/lECD). An Agilent HP-5 (crosslinked 5% PH ME siloxane) capillary column (30 m long, 0.25 mm internal diameter, and

0.25 l m film thickness) was used. Operating conditions were as follows:

injector 250 C, detector 350 C, oven temperature was held at 110 C for 5 min, then increased to 280 C at 14 C/min and hold 2 min. The carrier gas was nitrogen at a column flow rate of 1.6 mL/min. All samples were analyzed in duplicate. To determine the quality of the method, a recovery study was performed on ten replicates of blank meat mixture fat samples

and ten replicates of blank liquid culture media (initial contamination level lower than the detection limit) overspiked with PCBs. Mean recoveries ranged from 42.3 to 86.1 and 97.3 to 110.0% for liquid culture medium and meat mixture fat, respectively depending on the PCB congener. The limits of detection (ng mL 1 of culture medium) for liquid culture medium were 0.1 for PCBs 10, 28 and 153, 0.2 for PCB 52 and 138, and 0.4 for PCB

180. For meat mixture, the limits of detection (ng g 1 fat) were 0.2 for

PCBs 10, 28, and 52, and 0.5 for PCBs 138, 153, and 180.

2.3. Statistical analysis

Data were analyzed using the Statistica 6.0 software package. Results were analyzed by repeated measures analysis of variance (ANOVA): 2 culture media · 3 treatment · 11 incubation time for starter growth evalu- ations, and 2 culture media · 2 treatment · 11 incubation time for PCB degradation, except for results of PCB 10 that were evaluated by two-way ANOVA with no repeated measure due to the existence of groups without variance (all PCB levels lower than the limit of quantification). Post hoc analysis was carried out using Duncan’s test when appropriate. Differences were considered to be significant when p 6 0.05. The changes in PCB con-

centration over the incubation time were fitted using non-linear regression (SlideWrite Plus 4.0 software package) to an exponential equation:

y ¼ a þ b expð x=c Þ

3. Results

3.1. Effect of PCBs on the growth of meat starter

Addition of a mix of PCBs 10, 28, 52, 138, 153, and 180 (0.01 ppm of each compound) did not affect growth of S. xylosus in BHI or MSM (Fig. 2 ). In MSM counts increased

around 3 log 10 cfu/mL during the initial 12 h, while in BHI the increase was around 4 log 10 cfu/mL. This pattern was observed in the three treatments (control, acetone, and

mix of PCBs) with no significant differences among the

treatments.

3.2. In vitro degradation of PCBs by the meat starter

Staphylococcus xylosus

Figs. 3 and 4 summarize the percentage of PCBs remain-

ing in culture media after incubation with S. xylosus . These percentages are related to the initial concentration of PCBs (0 h). ANOVA did not reveal significant main effects or interactions between culture media and incubation time on the concentrations of PCB 10 (data not shown). Besides,

no significant exponential relationship was observed

between incubation time and the concentrations of PCB 10, indicating that this compound was not degraded up to 168 h of incubation regardless of the culture medium (data not shown). Although there were perceptible reductions in the con- centrations of PCB 28 (Fig. 3 A), 52 (Fig. 3 B), and 180 (Fig. 4 C), ANOVA revealed no significant main effects or interactions between culture media, treatment and

or interactions between culture media, treatment and Fig. 2. Growth rate of S. xylosus in (A)

Fig. 2. Growth rate of S. xylosus in (A) mineral salt medium (MSM) or (B) nutritive medium (BHI) during 168 h of incubation at 30 C. Medium were unspiked (control) or spiked with 0.01 ppm PCBs mix (congeners 10, 28, 52, 138, 153, and 180) or acetone (vehicle of PCBs mix). Results are the mean ± standard error of three independent experiments (standard errors were lower than the legend symbol).

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es et al. / Food and Chemical Toxicology 44 (2006) 847–854 Fig. 3. Degradation of PCB

Fig. 3. Degradation of PCB 28 (A) and PCB 52 (B) by the meat starter S. xylosus in culture media during incubation at 30 C. Liquid culture medium was spiked with 0.01 ppm of each PCB congener. The PCB concentrations found in the culture media not spiked with PCBs (control) were always lower than 10% and are omitted for clarity purposes. Results are expressed as percentage (mean ± standard error of three independent experiments) of PCB concentration at the beginning of the incubation (0 h).

incubation time on the concentrations of these congeners. This probably occurred due to fluctuations in the levels of these congeners during the experimental period. How- ever, regression analysis revealed highly significant expo- nential relationships between incubation time and the concentrations of PCB 28 ( r = 0.96, p < 0.05) and PCB 180 (r = 0.89, p < 0.05) in BHI (Figs. 3 A and 4 C). In MSM there was a highly significant exponential relation- ship between incubation time and the concentration of PCB 52 ( r = 0.84, p < 0.05) and 180 ( r = 0.97, p < 0.05), indicating that these compounds were signifi- cantly degraded during the incubation (Figs. 3 B and 4 C). ANOVA revealed a significant main effect of the incuba- tion time and a significant treatment · incubation time interaction for PCBs 138 and 153, indicating that these compounds were degraded both in BHI (78% and 68%, respectively) and in MSM (71% and 66%, respectively; Fig. 4 A and B). Maximum degradation occurred within 24 h for both compounds, which coincides with the loga- rithmic phase of growth of S. xylosus ( Fig. 2 ).

3.3. PCBs degradation in meat mixture

Fig. 5 shows the degradation of PCBs by the meat starter S. xylosus in meat mixture. The evolution of the concentrations during the incubation is expressed as percentage of PCB levels at the beginning of incubation. ANOVA did not reveal significant main effects or interac- tions between treatment and incubation time on the concentration of any compound evaluated. However,

regression analysis revealed highly significant exponential relationships between incubation time and PCB 10 (r = 0.88, p < 0.05), indicating that this compound was significantly degraded during incubation. No significant exponential relationship was observed between the incuba- tion time and the concentrations of the other PCB congen- ers evaluated (28, 52, 138, 153, and 180), indicating that these compounds were not significantly degraded in the meat mixture up to 168 h of incubation.

4. Discussion

4.1. Effect of PCBs on the growth of meat starter

The mix of PCBs 10, 28, 52, 138, 153, and 180 did not affect the growth of S. xylosus at 30 C during 24 h. In contrast, Bayarri (1997) observed a reduced growth of S. carnosus in two culture media (nutritive and mineral salt medium) containing a mix of organochlorine compounds and PCB 153. However, each microorganism has a dis- tinct behavior towards the stress caused by contaminants (Abraham et al., 2002 ) and no studies were found con- cerning the influence of PCBs on S. xylosus growth, which makes difficult the comparisons with our results. Besides, microbial population may adapt itself to the presence of organochlorine compounds by various mechanisms including the development of an appropriate enzymatic system to eliminate the stress factor, i.e. to degrade the contaminants of the medium ( Dolfing and Beurskens, 1995 ). This mechanism could explain the normal growth

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al. / Food and Chemical Toxicology 44 (2006) 847–854 851 Fig. 4. Degradation of PCB 138

Fig. 4. Degradation of PCB 138 (A), PCB 153 (B), and PCB 180 (C) by the meat starter S. xylosus in culture media during incubation at 30 C. Liquid culture medium was spiked with 0.01 ppm of each PCB congener. The PCB concentrations found in the culture media not spiked with PCBs (control) were always lower than 10% and are omitted for clarity purposes. Results are expressed as percentage (mean ± standard error of three independent experiments) of PCB concentration at the beginning of the incubation (0 h). Difference between the initial and final levels was statistically significa nt (Duncan’s test, p < 0.05) for PCB 138 and 153.

nt (Duncan’s test, p < 0.05) for PCB 138 and 153. Fig. 5. Degradation of PCBs

Fig. 5. Degradation of PCBs 10, 28, 52 (A), 138, 153 and 180 (B) in the meat mixture during incubation with S. xylosus. Meat mixture was spiked with 1 l g g 1 fat of each PCB congener and then stored at 30 C. Results (mean ± standard error of three independent experiments) are expressed as percentage of PCB concentration at the beginning of the incubation (0 h).

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of S. xylosus in the presence of PCBs when compared with the control.

4.2. In vitro degradation of PCBs by the meat starter S. xylosus

Microorganism metabolism may change depending on the environmental circumstances. Many factors like temper- ature, pH, and carbon content can strongly influence the rate, extent, and route of PCB degradation ( Wiegel and Wu, 2000; Borja et al., 2005 ). Microorganisms might prefer- entially degrade other organic substances before PCBs. This behavior can not be evaluated in mineral salts medium, where the only carbon source are the PCBs ( Katayama and Matsumura, 1993; Klasson et al., 1996 ). For this reason we investigated PCB degradation both in MSM and BHI. Accordingly, PCB 52 was significantly degraded in MSM, but not in BHI, where other carbon sources were available. In contrast, the degradation of the other congeners evalu- ated was similar in BHI and MSM medium. PCBs containing chlorine atoms in the para (4 and 4 0 )

and meta positions (3,3 0 ,5 and 5 0 ) are preferentially biode- graded ( WHO, 1993; Wiegel and Wu, 2000; Borja et al., 2005 ). Among the six PCB congeners evaluated only two had no chlorine atom in the para positions: PCB 10 (2,6- dichlorobiphenyl) and PCB 52 (2,2 0 ,5,5 0 -tetrachlorobiphe- nyl). Accordingly, PCB 10 was not significantly degraded both in MSM and BHI, while PCB 52 was degraded only in MSM. As discussed above the degradation of PCB 52 in MSM may have occurred because in this medium PCBs are the only carbon source, which may favor the attack to this compound that is not usually degraded. In our experimental design culture media were simulta- neously spiked with the six PCB congeners. Therefore, the transitory increase in the concentration of PCB 52 during incubation in BHI (although not statistically significant) could be attributed to the dechlorination of higher chlori-

congeners like PCBs 153 (2,2 0 ,4,4 0 ,5,5 0 -hexachlorobi-

phenyl) and/or 180 (2,2 0 ,3,4,4 0 ,5,5 0 -heptachlorobiphenyl).

This hypothesis is especially probable since the removal of their two chlorine atoms from the para (4,4 0 ) and meta (3) positions, which are preferential pathways for degrada- tion ( WHO, 1993; Wiegel and Wu, 2000; Borja et al., 2005 ), yields PCB 52. Degradation of these PCBs was sim- ilar both in MSM and BHI medium. Hence, the increase in the concentration of PCB 52 in BHI would depend on a reduced degradation of this compound in BHI. Fedi et al. (2001) evaluated PCB degradation by fifteen aerobic bacterial strains using biphenyl as the sole carbon and energy source and observed that some strains degraded up to 75% of PCB 28. Although we have observed a similar amount of PCB 28 degradation by S. xylosus , degradation was statistically significant only in BHI and not in MSM. Sierra et al. (2003) observed that after seven days of incu- bation in a mineral salt medium containing a commercial mixture of PCBs (Aroclor 1242), the new isolated aerobic bacterium Janibacter sp. degraded 100% of PCB 10 and

nated

92% of PCB 52. This report contrasts with our results where PCBs 10 and 28 were not significantly degraded by S. xylosus in MSM. These discrepancies suggest that the ability of microorganisms to degrade PCBs may be differ- entially affected by environmental conditions depending on the microorganism studied. Concerning to the degradation of PCBs 138, 153 and 180 in both culture media our results are in agreement with Bayarri (1997) , who verified that other microorganisms (Micrococcus varians and S. carnosus ) degrade highly chlo- rinated compounds, like PCB 153 (six chlorine atoms), in aerobic conditions. However, according to Abraham et al. (2002) , microorganisms generally degrade low chlori- nated PCBs in aerobic conditions, while highly chlorinated PCBs are commonly degraded in anaerobic conditions. Various authors have found microbial degradation of PCBs in aerobic conditions ( Ruiz-Aguilar et al., 2002; Komancova´ et al., 2003 ), but most studies evaluated micro- organisms isolated from soil, water or sediments. Few studies investigated the degradation of organochlorine com- pounds by starter microorganisms used in meat products. Mirna and Coretti (1979) observed that micrococci and lac- tobacilli obtained from a commercial starter culture degraded lindane (70% and 90%, respectively) after two weeks of incubation in nutritive medium. DDT was inten- sely degraded and converted into DDD by micrococci, but was not degraded by lactobacilli. Bayarri et al. (1998) observed that some organochlorine compounds were degraded by the meat starter microorganism M. varians in an in vitro study in mineral salt medium. Although these studies have not evaluated PCB degradation, results found were similar to those that we obtained with S. xylosus and indicate that starter microorganisms are able to degrade some chlorinated compounds. This degradation could be important in the reduction of the toxicological risk assoc- iated with residues of PCBs in meat products.

4.3. PCBs degradation in meat mixture

Many authors investigated the effect of aerobic and anaerobic microorganisms on PCBs levels in organic matrixes, especially in soil and sediments ( Chang et al., 2001; Fedi et al., 2001; Nawab et al., 2003; Sierra et al., 2003 ). A low number of investigations using meat as organic matrix were performed ( Mirna and Coretti, 1979; Arin˜ o et al., 1993; Bayarri et al., 1998; Abou-Arab, 2002 ), and in these experiments various starter microorgan- isms were used. However, as far as we know no study has reported the ability of S. xylosus to degrade PCBs. According to Arin˜ o et al. (1995) microorganisms used in meat products enzymatically degrade part of the organo- chlorine compounds, by using these compounds as carbon sources. This degradation could be favored by many fac- tors allowing the enzyme–substrate contact, like the mince process and the mixture of the ingredients. Spatial disposition of the halogenated substitutions influences PCB degradation ( Bedard et al., 1986; Fetzner

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and Lingens, 1994 ). Bopp (1986) found that ortho -substi- tuted PCB congeners are highly resistant to degradation by Pseudomonas strain LB400. In contrast, in the present study, where PCBs 10, 52, 138, 153, and 180 are di- ortho congeners, we observed a reduction in the levels of PCB 10 in a meat mixture. The resistance of PCB 52 to degrada- tion is of concern, since this PCB has been associated to malignant lesions ( Lucena et al., 2001 ). Degradation of non- ortho and mono-ortho substituted congeners are also very important, because these compounds are structurally related to 2,3,7,8-tetrachlorodibenzo-p- dioxin and have dioxin-like effects, i.e. carcinogenic effects ( Penteado and Moreira Vaz, 2001 ). The mono- ortho conge- ner evaluated in the present study (PCB 28) was significantly degraded only in the BHI medium. Some authors indicate that degradation magnitude is inversely related to the number of chlorine atoms in the compound. Accordingly, the dichloro-substituted congener PCB 10 was the only PCB significantly degraded during meat mixture incubation, while higher chlorinated PCBs were slightly and non-significantly degraded. Similar to our study, Bayarri (1997) did not observe sig- nificant reduction in the levels of PCB 153 during sausage maturation with commercial starter microorganisms con- taining Pediococcus acidilactici , Pediococcus pentosaceus, and M. varians . Abou-Arab (2002) investigated the ability of two strains of meat starter microorganisms ( Lactobacillus plantarum and M. varians ) to degrade organochlorine pesticides in sausage mixture. After 72 h it was observed a reduction of 10% in p ,p 0 -DDT levels and 18% in lindane levels. Arin˜ o et al. (1993) found a 30% reduction in the lindane levels in sausage after 30 days of curing, indicating the possible action of the fermented meat microflora.

5. Conclusion

In summary, it was shown that although S. xylosus decreased the concentration of various PCB congeners in liquid media, it was less effective in meat mixture. It can be inferred that the use of the starter microorganism S. xylosus in meat products would have a limited contribu- tion to eliminate PCB residues, since only PCB 10 was sig- nificantly degraded in the meat mixture.

Acknowledgement

This work was supported by CNPq (Conselho Nacional de Desenvolvimento e Pesquisa). I. Costabeber was the recipient of PROFIX Fellowship. T. Emanuelli is the recipient of a CNPq Research Fellowship.

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